CN105784646A - Anti-background-interference simultaneous extraction method of enhanced image of multi-target precious metal nano particle pair - Google Patents
Anti-background-interference simultaneous extraction method of enhanced image of multi-target precious metal nano particle pair Download PDFInfo
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Abstract
The invention discloses an anti-background-interference simultaneous extraction method of an enhanced image of multi-target precious metal nano particle pairs. Aiming at a complex biological environment in which macromolecular groups exist, by using a resonant coupling effect of surface plasmas of the precious metal nano particle pairs, the invention discloses a method which can realize optimal excitation conditions of the precious metal nano particle pairs which are randomly distributed in directions and simultaneously match and remove macromolecular background noises. The method can realize no target loss in the biological environment with macromolecular group noise interference and development of all precious metal nano particle pairs which are randomly distributed in directions.
Description
Technical field
The invention belongs to the single molecular recognition in life science and nano-probe technical field, relating to the detection of metallic nano detecting probe pair, to be related specifically in tested surface sparse is scattered and the best is strengthened acquisition methods while amplitude image picture by multiple nano metal probes that orientation is random.
Background technology
There is the noble metal nano particles of surface plasmon resonance effect, for instance golden nanometer particle, there is high brightness, without the feature such as photobleaching, bio-compatibility be good, thus become the label of a kind of probe biomolecule received much concern.Moreover, they have the surface plasmon resonance effect of uniqueness, extremely strong absorption is shown at visible near infrared band, especially when two particles are close together generation surface plasma body resonant vibration coupling, gap location at this particle pair produces the electric field strengthened, and the delustring intensity of comparable single particle increases several times.Therefore using of noble metal nano particles pair has important being worth and wide application space in strengthening molecular image image and bio-sensing sensitivity etc..But this potentiation is but strongly depend on shooting condition, the polarization state requiring exciting light needs with this axle center that line of centres of particle is constituted to be mated and could obtain the best reinforced effects, such as it is shown in figure 1, when the polarization state of exciting light and the axis of particle pair are parallel, strengthen maximum;When both are vertical, reinforced effects is minimum.The test of testing sample can exist multiple particle pair simultaneously, the spatial arrangements of the axis of every pair of particle and position distribution are all random, if only making partial target therein obtain coupling, and its image enhaucament is displayed, other target image not obtaining enhancing can be submerged and lose, it is therefore desirable to the particle of all random alignment to all carrying out enhancing condition coupling.In addition, having there is also biomacromolecule group in biological sample, the dominant foot in their size is to flood the signal of noble metal nano particles pair.Therefore in the biotic environment having macromolecular mass to disturb driftlessness lose detect noble metal nano particles to being by the key that quantitative biochemical is analyzed, the proposition of the present invention can realize multi-target particle in the biotic environment having macromolecular mass disturb and the Auto-matching of shooting condition and strengthening without the best to losing of particle is developed while image.
Summary of the invention
The technical problem to be solved in the present invention is automatic shooting condition coupling and the acquisition methods simultaneously of the best enhancing image of the multiple nano metal probes pair realizing tested surface inner orientation random alignment in the biotic environment having macromolecular mass to disturb.
The technical scheme is that
A kind of not by the extraction method strengthening image of the multiple target noble metal nano particles pair of ambient interferences, it is characterized in that noble metal nano particles is the metal nanoparticle with surface plasmon resonance effect, the axis and the incident illumination that change noble metal nano particles pair in tested plane continuously excite the angle between polarization state so that the axis of every a pair noble metal in tested plane obtains equal polarization state MM meeting in certainty of measurement.Specifically include following steps:
Step 1: set the wavelength of incident illumination as the noble metal nano particles surface plasma body resonant vibration coupled wavelength to producing;
Step 2: the polarization state of incident illumination projecting direction in tested plane is P, rotates the direction half cycle of P or the integral multiple of half cycle continuously with the step-length set;Often rotate a step-length, measure and record a width amplitude image picture;After completing all rotations, it is thus achieved that n width amplitude image picture;
Step 3: n width image step 2 obtained resets overlapping about tested plane coordinate system;For the same location point in tested plane, all amplitude image pictures being carried out original position comparison, if change rate of strength is less than the threshold value arranged, then this test point is judged as impurity, and is rejected;
Step 4: repeat step 3, carries out impurity to each point in tested plane and judges and reject;
Step 5: all amplitude image pictures after eliminating impurity are carried out original position superposition, the image generated after superposition is the enhancing image that every pair of noble metal nano particles all obtains that incident light polarization state mates with its axis;
The setting principle of described step-length is: span is 0 to 45 degree, and it is more little that step-length is arranged, and the accuracy of identification of target is more high, measures the spent time more long;
The change rate of strength of described amplitude image picture be same measured point maximum in n width image divided by
Minima;The span of the change rate of strength threshold value of amplitude image picture is be more than or equal to 2.
The effect of the present invention is can highly sensitive realization by the noble metal nano particles multi-targets recognition to labelling and to develop in the complex biological environment having macromole to exist simultaneously.
Accompanying drawing explanation
Accompanying drawing 1 be between particle coupled electric field amplitudes with incident ray polarization direction and the particle variation relation simulation result figure to the angle of the line of centres.In figure: the particle pair that centre of sphere spacing is 65nm that two gold particle of diameter respectively 60nm are constituted;X-axis is incident ray polarization direction and the particle angle (unit: degree) to the line of centres;Y-axis is double; two particle coupled electric field amplitudes is normalization when 90 degree relative to X.
Accompanying drawing 2 is that single noble metal nano particles, biomacromolecule group, the noble metal test sample to coexisting strengthen change schematic diagram when incident ray polarized light different directions angle.
In figure: 1 single noble metal nano particles;2 noble metals pair;
3 and 4: biomacromolecule group;5 are present in the particle enhancing electric field to gap location;
Represent the polarization state direction of incident illumination.
Detailed description of the invention
Accompanying drawing 2 is that single noble metal nano particles, biomacromolecule, the noble metal test sample to coexisting strengthen change schematic diagram when incident light polarization direction difference, the polarization state of incident illumination according to being rotated clockwise, illustrates 4 position of rotation to be 0 degree the be starting point of the X-axis angle with tested plane coordinate system in figure.For a pair gold goal, the diameter of two gold goals respectively 60nm, centre of sphere spacing is 65nm, as shown in Figure 1, along with the change of incident ray polarization direction in the process rotated, the surface plasma body resonant vibration coupling of double; two particle gap locations strengthens intensity and strong change occurs.When polarization state is parallel with double; two particle centre lines, coupling strengthens maximum;And when polarization state is perpendicular to double; two particle centre line, coupling strengthens disappearance.Single metal nanoparticle and biomacromolecule group do not have surface plasma body resonant vibration coupling enhancement effect, and in rotary course, the rate of change of its oscillator intensity is much smaller than the rate of change of the oscillator intensity of bimetal nano particles.A kind of not by the extraction method strengthening image of the multiple target noble metal nano particles pair of ambient interferences, it is characterized in that noble metal nano particles is the metal nanoparticle with surface plasmon resonance effect, the axis and the incident illumination that change noble metal nano particles pair in tested plane continuously excite the angle between polarization state, the axis making every a pair noble metal in tested plane obtains equal polarization state MM meeting in certainty of measurement, comprises the steps:
Step 1: set the wavelength of incident illumination as the noble metal nano particles surface plasma body resonant vibration coupled wavelength to producing;
Step 2: the polarization state of incident illumination projecting direction in tested plane is P, rotates the direction half cycle of P or the integral multiple of half cycle continuously with the step-length set;Often rotate a step-length, measure and record a width amplitude image picture;After completing all rotations, it is thus achieved that n width amplitude image picture;
Step 3: n width image step 2 obtained resets overlapping about tested plane coordinate system;For the same location point in tested plane, all amplitude image pictures being carried out original position comparison, if change rate of strength is less than the threshold value arranged, then this test point is judged as impurity, and is rejected;
Step 4: repeat step 3, carries out impurity to each point in tested plane and judges and reject;
Step 5: all amplitude image pictures after eliminating impurity are carried out original position superposition, the image generated after superposition is the enhancing image that every pair of noble metal nano particles all obtains that incident light polarization state mates with its axis;
A kind of not by the extraction method strengthening image of the multiple target noble metal nano particles pair of ambient interferences, it is characterized in that the setting principle of step-length is: span is 0 to 45 degree, it is more little that step-length is arranged, and the accuracy of identification of target is more high, measures the spent time more long;
A kind of not by the extraction method strengthening image of the multiple target noble metal nano particles pair of ambient interferences, its spy is the change rate of strength of amplitude image picture is that same measured point maximum in n width image is divided by minima;
A kind of not by the extraction method strengthening image of the multiple target noble metal nano particles pair of ambient interferences, its spy is that the span of the change rate of strength threshold value of amplitude image picture is for be more than or equal to 2.
Claims (4)
1. the extraction method strengthening image of the multiple target noble metal nano particles pair not being subject to ambient interferences, it is characterized in that noble metal nano particles is the metal nanoparticle with surface plasmon resonance effect, the axis and the incident illumination that change noble metal nano particles pair in tested plane continuously excite the angle between polarization state, the axis making every a pair noble metal in tested plane obtains equal polarization state MM meeting in certainty of measurement, comprises the steps:
Step 1: set the wavelength of incident illumination as the noble metal nano particles surface plasma body resonant vibration coupled wavelength to producing;
Step 2: the polarization state of incident illumination projecting direction in tested plane is P, rotates the direction half cycle of P or the integral multiple of half cycle continuously with the step-length set;Often rotate a step-length, measure and record a width amplitude image picture;After completing all rotations, it is thus achieved that n width amplitude image picture;
Step 3: n width image step 2 obtained resets overlapping about tested plane coordinate system;For the same location point in tested plane, all amplitude image pictures being carried out original position comparison, if change rate of strength is less than the threshold value arranged, then this test point is judged as impurity, and is rejected;
Step 4: repeat step 3, carries out impurity to each point in tested plane and judges and reject;
Step 5: all amplitude image pictures after eliminating impurity are carried out original position superposition, the image generated after superposition is the enhancing image that every pair of noble metal nano particles all obtains that incident light polarization state mates with its axis.
2. a kind of not by the extraction method strengthening image of the multiple target noble metal nano particles pair of ambient interferences as claimed in claim 1, it is characterized in that the setting principle of step-length is: span is 0 to 45 degree.
3. a kind of not by the extraction method strengthening image of the multiple target noble metal nano particles pair of ambient interferences as claimed in claim 1 or 2, its spy is the change rate of strength of amplitude image picture is that same measured point maximum in n width image is divided by minima.
4. a kind of not by the extraction method strengthening image of the multiple target noble metal nano particles pair of ambient interferences as claimed in claim 3, its spy is that the span of the change rate of strength threshold value of amplitude image picture is for be more than or equal to 2.
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CN107101942A (en) * | 2017-05-17 | 2017-08-29 | 大连理工大学 | A kind of probe for being used to polarize micro-imaging based on bimetal nano particles |
CN108414423A (en) * | 2018-04-27 | 2018-08-17 | 成都理工大学 | The scanning electron microscope imaging method of the micro-nano hole of rock |
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CN102539400A (en) * | 2011-12-31 | 2012-07-04 | 广东工业大学 | High-accuracy fluorescence anisotropy microscopic imaging device and method |
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CN108414423A (en) * | 2018-04-27 | 2018-08-17 | 成都理工大学 | The scanning electron microscope imaging method of the micro-nano hole of rock |
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